Cloning and Expression of Nanyang Cattle BoLA-DRA Gene in Escherichia coli

This study aimed to clone Nanyang cattle BoLA-DRA gene,construct recombinant prokaryotic expression vector and to study the gene expression in Escherichia coli.Firstly,BoLA-DRA gene fragment was amplified by RT-PCR after total RNA was extracted from spleen of Nanyang cattle.Following,the BoLA-DRA gene was cloned into the pGEM-T vector and transferred into E.coli DH5α.After sequence analysis,the BoLA-DRA gene was cloned into prokaryotic expression vector pGEX-4T-1 and induced by IPTG(isopropyl β-D-1-thiogalactopyranoside).Results showed that the 917 bp long BoLA-DRA gene was cloned,the fusion protein was obtained successfully,and it existed mostly in the form of inclusion body.Identification of expressed products by SDS-PAGE and Western blot showed that the target protein was 54.4 ku which was consistent with the expected results.The results laid a foundation for further studying on the function of the protein and preparation of antibody.